title: Construction and characterization of a novel chimeric antibody c3C7 specific for the integrin αIIbβ3 complex.
abstract: A murine monoclonal antibody (mAb) 3C7 against integrin αIIbβ3 was previously obtained as a potential antithrombotic agent in our laboratory. The epitope of 3C7 is a specific conformation of the αIIbβ3 complex, but not either of the two subunits, which makes it different from abciximab, a supplementary antibody drug used in percutaneous coronary intervention which has a cross-reaction with other integrins sharing the β3 subunit. To reduce the human anti-mouse antibody reactions of 3C7, the variable regions of this antibody were cloned and fused with the constant counterparts of human IgG1. Two vectors of light and heavy chains were constructed and co-transfected into CHO-dhfr(-) cells. The chimeric antibody c3C7 was purified and the properties of c3C7 were compared with 3C7. Identical to its parent antibody 3C7, c3C7 binds to the αIIbβ3 complex, but not to either of the subunits. The K(d) value of c3C7 was in the same order of magnitude as 3C7 (1.570 ± 0.326 vs 0.780 ± 0.182 nmol/L). Human platelet aggregation induced by adenosine diphosphate was effectively inhibited by c3C7 in a dose-dependent manner. In conclusion, after the modification, c3C7 retained the properties of its parent mAb with no loss of its biological activity. Therefore, c3C7 has the potential to become a novel agent for the treatment of thrombosis.
authors.affiliation: Institute of Molecular Medicine, Nanjing University, Nanjing, 210093, China, jianaq@nju.edu.cn.
authors.forename: Aiqin
authors.initials: A
authors.lastname: Jiang
authors.forename: Wang
authors.initials: W
authors.lastname: Zhang
authors.forename: Qiong
authors.initials: Q
authors.lastname: Wu
authors.forename: Wenbo
authors.initials: W
authors.lastname: Jin
authors.forename: Yanchun
authors.initials: Y
authors.lastname: Tang
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
authors.forename: Jian-Ning
authors.initials: JN
authors.lastname: Liu
mesh_terms: D000818:Animals; D000906:Antibodies / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D006801:Humans; D007074:Immunoglobulin G / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D051379:Mice; D010974:Platelet Aggregation / Q000187:drug effects; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D000906:Antibodies; D007074:Immunoglobulin G; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D011994:Recombinant Proteins